Patch Compartment Research Articles

Colocalization of serotonin receptor subtypes 5-HT2A, 5-HT2C, and 5-HT6 with neuropeptides in rat striatum.

There are two primary output pathways from the striatum: a projection to the globus pallidus, and projection to the substantia nigra. Certain striatally expressed neuropeptides are differentially distributed between these two pathways. Specifically, enkephalin is expressed in striatopallidal neurons, whereas substance P and dynorphin are expressed in striatonigral neurons. Several serotonin receptors are also prominently expressed in the striatum, but little is known about how they fit into the molecular neuroanatomy described above. We used double-label in situ hybridization to determine the striatal distribution of the mRNAs of the serotonin2A (5-HT2A), serotonin2C (5-HT2C), and serotonin6 (5-HT6) receptors in relation to enkephalin, substance P, and dynorphin expressing output neurons. Rat brain sections were simultaneously hybridized with an 35S riboprobe for one of the serotonin receptors and a digoxygenin labeled riboprobe for one of the neuropeptides. Sections were examined by using brightfield microscopy, and the degree of colocalization of the two mRNAs determined. All the serotonin receptors colocalized extensively with all three of the neuropeptides examined. None of the serotonin receptors showed preferential colocalization in striatopallidal (enkephalin containing), or striatonigral (substance P or dynorphin containing) cells. The 5-HT2A and 5-HT2C mRNAs displayed a differential distribution with regard to the scattered islands of strongly dynorphin mRNA positive cells, which are thought to reside in the striatal patch compartment. Within these islands, 5-HT2C mRNA expression was much higher than in surrounding areas. 5-HT2A mRNA showed the opposite pattern with decreased expression over dynorphin rich cell clusters.

Differential messenger RNA expression of prodynorphin and proenkephalin in the human brain.

The opiate system is involved in a wide variety of neural functions including pain perception, neuroendocrine regulation, memory, drug reward, and tolerance. Such functions imply that endogenous opioid peptides should have anatomical interactions with limbic brain structures believed to be involved in the experience and expression of emotion. Using in situ hybridization histochemistry, the messenger RNA expression of the opioid precursors, prodynorphin and proenkephalin, was studied in whole hemisphere human brain tissue. Different components of the limbic system were found to be characterized by a high gene expression of either prodynorphin or proenkephalin messenger RNA. Brain regions traditionally included within the limbic system (e.g. amygdala, hippocampus, entorhinal cortex and cingulate cortex) as well as limbic-associated regions including the ventromedial prefrontal cortex and patch compartment of the neostriatum showed high prodynorphin messenger RNA expression. In contrast, high levels of proenkephalin messenger RNA were more widely expressed in the hypothalamus, periaqueductal gray, various mesencephalic nuclei, bed nucleus of the stria terminalis, and ventral pallidum; brain regions associated with endocrine-reticular-motor continuum of the limbic system. The marked anatomical dissociation between the expression of these two opioid peptide genes, seen clearly in whole hemisphere sections, indicates that distinct functions must be subserved by the prodynorphin and proenkephalin systems in the human brain.

Differential maturation of cholinergic interneurons in the striatal patch versus matrix compartments

Differential maturation of cholinergic interneurons in the striatal patch versus matrix compartments

Somatostatin and the patch/matrix compartments of the rat caudate-putamen.

In the caudate-putamen of the rat, two subpopulations of medium aspiny neurons exist that contain somatostatin. The first subpopulation contains somatostatin 14, somatostatin 28, and somatostatin 28(1-12). The other subpopulation contains only somatostatin 28. To examine the relationship between somatostatin-containing neurons and the patch/matrix compartments, a series of double-labelling experiments using antisera directed against different somatostatin peptides and calbindin were used. Sections stained in this manner were examined with the aid of a confocal microscope. The results of these experiments indicate that somatostatin 28(1-12)-containing neurons may play a role in matrix integration, with some input directed from the patch compartment. In addition, somatostatin 28-containing neurons are more numerous in the patch compartment than somatostatin 28(1-12)-containing neurons, suggesting a possible role for these neurons in patch integration.

Patterns of convergence and segregation in the medial nucleus accumbens of the rat: relationships of prefrontal cortical, midline thalamic, and basal amygdaloid afferents.

In the rat, fibers from the prelimbic cortex terminate in the medial nucleus accumbens. Anterior paraventricular thalamic and parvicellular basal amygdaloid fibers reached both the prelimbic cortex and the medial nucleus accumbens. All three afferent systems have an inhomogenous distribution within the nucleus accumbens, and whether or not these projections actually reach the same areas is unknown. Our aim was to evaluate the relationships of the three afferents with respect to the shell, the core, and the cell clusters of the nucleus accumbens. Double anterograde tracing and single anterograde tracing combined with immunohistochemistry for calbindin (D28k) or Nissl stain was used. Following tracer injections in the prelimbic cortex and the anterior paraventricular thalamus, a complementary (i.e., nonoverlapping) pattern of fibers was found in the shell. Thus, afferents from the prelimbic cortex are associated with cell clusters, whereas those from the anterior paraventricular thalamus avoid these cells but are affiliated with regions exhibiting weak homogeneous calbindin immunoreactivity. In the calbindin-poor patches of the core, the situation is reversed as both sets of fibers overlap. In cases with injections in the prelimbic cortex and the parvicellular basal amygdala, a pattern of overlap was seen in the shell and core. Thus, the fibers in the shell were found together in association with cell clusters, whereas regions of weak homogeneous calbindin immunoreactivity were avoided. In the core, overlap was seen in the patch compartment. Finally, with parvicellular basal amygdala/paraventricular thalamus injections, a complementary fiber organization was present in the shell, but overlap was prominent in the patches of the core. The results demonstrate that the relationships of prelimbic cortical, paraventricular thalamic, and parvicellular basal amygdaloid afferents in the nucleus accumbens vary according to their compartmental (immunohistochemical and cellular) affiliation. Compartmentalization is therefore a possible anatomical substrate for condensation or segregation of neuronal signals passing through the nucleus accumbens.

Pattern Formation in the Mammalian Forebrain: Striatal Patch and Matrix Neurons Intermix Prior to Compartment Formation

The striatum of the mammalian forebrain is divided into two compartments: the patches and the matrix. Neurons of the patch compartment in the rat striatum become postmitotic earlier in neurogenesis than neurons of the matrix compartment. The selective adhesion of patch neurons to one another has been suggested previously to be an important developmental mechanism of striatal compartmentation. We asked if the selective adhesion of patch neurons is expressed before or after the migration of the majority of the matrix neurons into the striatum. Patch neurons were labelled in vivo by a fluorescent retrograde tracer injected into the substantia nigra on embryonic day 19, which almost exclusively labelled patch neurons. Matrix neurons were labelled with a maternal injection of bromodeoxyuridine at embryonic day 18. When animals were killed at embryonic day 20, the majority of the retrogradely labelled patch neurons were intermixed with the bromodeoxyuridine-labelled matrix neurons, although there appeared to be clustering of some of the patch neurons. However, by postnatal day 2 there was a complete segregation of the clusters of the retrogradely labelled patch neurons from the bromodeoxyuridine-labelled matrix neurons in the striatum. This process was modelled in vitro. The patch and matrix compartments were labelled in vivo at embryonic day 13 and 18 respectively, with different birthdate markers ([3H]thymidine or bromodeoxyuridine). At embryonic day 20 the striatal tissue was removed, dissociated and reaggregated in suspension cultures. After 1 day in vitro, labelled patch and matrix neurons were randomly intermixed within the reaggregates. Examination of the cultures at 2.5 and 4 days in vitro revealed clumping of the labelled patch neurons towards the centres of the reaggregates. Over this same period, the labelled matrix neurons did not clump and were dispersed towards the periphery of the reaggregates. The results suggest that patch neuron adhesiveness may appear relatively soon after these neurons become postmitotic, but that this adhesiveness is unable to overcome the initial force produced by the massive migration of matrix neurons into the striatum. We hypothesize that a migratory phase of embryonic striatal development exists, when fated patch and matrix neurons intermix. After this migratory phase, patch neuron adhesiveness can produce the mature segregation of the striatal compartments.

Ontogeny of the D1 Dopamine Receptor in the Rat Striatonigral System: an lmmunohistochemical Study

Antibodies were raised against a recombinant protein to analyse the pre- and postnatal ontogeny of the neurons expressing the D1 dopamine receptor in the striatum by immunohistochemistry. We report that D1 immunoreactivity is detectable from gestational day (G) 15 and is distributed homogeneously throughout the striatum from G15 to G18. From G19-20 to postnatal day (P) 3, D1 immunoreactivity becomes heterogeneous and predominates in cell bodies of the patch compartment while very limited immunoreactivity is detectable in the matricial compartment. The differential intensity between patches and matrix reaches its peak around P0. From P2, the pattern of D1 immunoreactivity progressively assumes the homogeneous distribution characteristic of the adult striatum. The expression of D1 mRNA in striatal neurons, as investigated by in situ hybridization, displays a similar pattern during this period. Substance P mRNA is also preferentially expressed in the patch compartment during the same period. D1 immunoreactivity appears at G17 in the substantia nigra as clusters of fibres and increases subsequently until reaching its adult form during the first postnatal week. These results demonstrate that the two compartments of the developing striatum display differential transcriptional and translational activity for the D1 gene and consequently two different and successive patterns of expression of D1 protein: patch neurons first express D1 receptor intensely while matrix neurons express it later and in smaller amounts so that D1 receptor appears transiently during the perinatal period as a marker of the patch compartment.

The human neostriatum shows compartmentalization of neuropeptide gene expression in dorsal and ventral regions: an in situ hybridization histochemical analysis.

Expression of neuropeptide messenger RNAs in striatal neurons was studied in post mortem human brain tissue by the use of in situ hybridization histochemistry. Clusters of cells expressing high levels of prodynorphin messenger RNA, and less strikingly, preprotachykinin messenger RNA, were prominent in the caudate nucleus and were present but less pronounced in the putamen. Proenkephalin and prosomatostatin messenger RNA-containing cells were more homogeneously distributed throughout the striatum, though the latter were much sparser. The four neuropeptide messenger RNA patterns in the nucleus accumbens were rather homogeneous compared with the dorsal striatum. Of these, prodynorphin messenger RNA showed a higher level of expression per cell in the nucleus accumbens relative to the dorsal striatum. The relationship of neuropeptide-containing cell clusters to the striosomal organization was characterized by looking at the register of these markers with patterns of low acetylcholinesterase activity and dense mu opiate receptor binding. In the caudate and putamen, clusters of cells expressing high levels of dynorphin and preprotachykinin messenger RNAs were clearly in register with the striosomes. The accumbens was defined by high prodynorphin messenger RNA levels, both low and high levels of acetylcholinesterase staining, and very low to absent mu opiate receptor binding. The distribution of high-expressing prodynorphin messenger RNA-containing cells--to the patch compartment and throughout the entire ventral striatum/nucleus accumbens region--defines the limbic domain of the neostriatum and suggests particular relevance to human striatal organization and function, because the distribution of this opioid neuropeptide is considerably more compartmentalized in human than in non-human species.

PET imaging of neuroreceptors in schizophrenia

Among the brain imaging techniques developed during the past two decades positron emission tomography has the highest sensitivity, allowing the analysis of specific neurotransmitter mechanisms in the living human brain. By using a combination of selective ligands labelled with positron emitting isotopes, D 1 and D 2 dopamine, serotonin 5HT 2 and benzodiazepine receptors were examined in schizophrenic patients (DSM-IIIR) and healthy control subjects. With this technique receptor populations could be excellently visualized and quantified with regard to number and binding characteristics in several brain regions. The characteristics of total D 1 and D 2 dopamine receptor populations in the caudate and putamen did not differ significantly in young drug naive schizophrenic patients and age matched control subjects. On the other hand, there was a highly significant reduction of the D 1 signal in high intensity regions of the basal ganglia when [ 11C]SCH 23390, a selective D 1 dopamine receptor antagonist, was used. These results suggest the possibility of a reduced D 1 dopamine receptor density in the patch compartment of the basal ganglia in schizophrenia. For 5HT 2 and benzodiazepine receptors no major alteration of receptor characteristics was observed in several neocortical and limbic brain regions.

Genesis and migration patterns of neurons forming the patch and matrix compartments of the rat striatum

The mammalian striatum is divided into two compartments, the patch (or striosome) and the matrix, which differ on the basis of several cytochemical markers, connection patterns, and time of neurogenesis. In the rat, the patch compartment consists of clusters of neurons isolated by matrix neurons; included in the patch compartment is a rim of neurons subjacent to the corpus callosum and external capsule, called the subcallosal streak. To study the genesis and migration patterns of striatal neurons forming these compartments, we injected pregnant rats with 5-bromo-2′-deoxyuridine (BrdU, which is incorporated into DNA during S-phase mitosis) on embryonic (E) day 14, to label patch neurons, or on E19, to label matrix neurons. Embryos were sacrificed at intervals after injection, for detection of BrdU by immunocytochemistry. Cells labeled at E14 were distributed fairly uniformly in the differentiated portion of the caudate-putamen through E19. However, by the day of birth (P0), E14-labeled cells were clustered into patches and the subcallosal streak. Using double immunocytochemistry for BrdU and for the patch marker substance P, we demonstrated a caudal-rostral gradient in the birth dates of neurons in the patch compartment; E14-labeled cells occupied substance P-labeled patches at the level of the posterior limb of the anterior commissure, but patches further rostral were nearly devoid of E14-labeled cells. The distance between the lateral ventricle and the nearest E14-labeled cells was greater on E19 than on E16 or on P0, suggesting secondary movement of early-born neurons during the process of cluster formation. Neurons labeled at E19 formed the matrix surrounding clusters of unlabeled cells, except in the nucleus accumbens (ventral striatum), where E19-labeled cells formed clusters. The data suggest that the uniformly-distributed population of early-born neurons is disrupted by the invasion of later-born (matrix) neurons, forcing the early-born neurons into clusters which are displaced toward the ventricular surface to form the patch compartment. Early-born neurons adjacent to the external capsule are not displaced, forming the subcallosal streak.

Irreversible blockade of D2 dopamine receptors by fluphenazine-N-mustard increases D2 dopamine receptor mRNA and proenkephalin mRNA and decreases D1 dopamine receptor mRNA and mu and delta opioid receptors in rat striatum.

The consequences of irreversibly-inhibiting D2 dopaminergic receptors on the expression of D1 and D2 dopamine receptor mRNAs and proenkephalin mRNA and on the levels of mu- and delta-opioid receptors in rat striatum were studied following single or repeated administration of the irreversibly-acting D2 dopamine receptor antagonist, fluphenazine-N-mustard (FNM). The density of dopamine and opioid receptors was determined by receptor autoradiography and the levels of the mRNA for the D1 and D2 dopamine receptors and proenkephalin were measured by in situ hybridization histochemistry. Repeated treatment of rats with FNM for 6 days produced more than 80% inhibition of D2 dopamine receptors but less than 25% inhibition of D1 dopamine receptors. Repeated treatment with FNM also resulted in statistically significant increases in D2 dopamine receptor mRNA but decreases in D1 dopamine receptor mRNA. In contrast, acute treatment with FNM for 3 h had no significant effects on D1 or D2 dopamine receptor mRNAs in striatum. An examination of the effects of FNM on the opioid system showed that repeated treatment with FNM for 6 days produced more than a 2-fold increase in the expression of proenkephalin mRNA in striatum. This was accompanied by significant decreases in mu- and delta-opioid receptors in striatum, mainly by reducing the size of the patch compartment of striatum. Acute treatment with FNM for 3 h produced small increases in proenkephalin mRNA and mu-opioid receptors in striatum but had no significant effects on delta-opioid receptors. These results suggest that persistent inhibition of D2 dopamine receptors differentially regulates the expression of D1 and D2 dopamine receptor mRNA in striatum, and that the magnitude, duration and interval of inhibiting dopaminergic transmission may be important factors in regulating dopamine receptor mRNA expression. These results also suggest that D2 dopamine antagonists indirectly down-regulate opioid receptors by increasing the expression of proenkephalin mRNA, thereby increasing enkephalin which, in turn, decreases opioid receptors in striatum.

Identification and characterization of striatal cell subtypes using in vivo intracellular recording and dye‐labeling in rats. III. Morphological correlates and compartmental localization

In the first two reports of this series, in vivo intracellular recording techniques were used to characterize the electrophysiological properties of two types of striatal neurons that had been identified by their distinct response patterns to stimulation of corticostriatal afferents. In this paper, we examined whether cells showing Type I or Type II response patterns also differed with respect to their morphology or compartmental localization by combining intracellular recording and Lucifer yellow staining with immunocytochemical localization of calbindin 28 kd immunoreactivity. In the majority of cases, both Type I and Type II neurons exhibited similar morphological characteristics, with 80% of the Type I cells (13/16) and all of the Type II cells (n = 40) being small or medium spiny neurons. In each case where the morphological class of the cell was different than the spiny cell class, the cell exhibited a Type I response pattern. These spiny neurons had somata that averaged 17.1 +/- 1.3 microns in diameter and gave rise to between four and eight primary dendrites. The axons typically arose from cell bodies (7/13 for Type I and 25/40 for Type II cells) and emitted extensive local axonal collaterals. However, the axons of Type I cells more frequently originated from the dorsal surface of the somata (9/13; 69%), whereas Type II axons more frequently arose from the ventral surface of the somata (25/35; 71%), which may account for their different extracellular waveforms. In coronally sectioned tissue (n = 18), the axons always projected laterally when the somata were located in the medial striatum and projected medially when the somata were in the lateral striatal region. In a subset of experiments (N = 22), Lucifer yellow-stained neurons were localized with respect to their position within the patch and matrix compartments of the striatum using subsequent staining for calbindin 28 kd immunoreactivity. Of the 20 labeled medium spiny neurons examined (Type II: N = 13; Type I: N = 7), 19 were located in the calbindin-positive matrix compartment. The only neuron localized to the patch compartment was a medium spiny cell that exhibited a Type II paired impulse response pattern. In addition, of the two aspiny neurons from this group with beaded dendrites, one was localized to the border between adjacent patch and matrix compartments, whereas the other was located completely within the matrix compartment. Therefore, despite their distinct paired impulse response patterns, the majority of both Type I and Type II neurons were medium spiny cells located in the matrix compartment of the striatum.(ABSTRACT TRUNCATED AT 400 WORDS)

Spatial organization of patch and matrix compartments in the rat striatum

The visualization of mu opiate receptors by [ 3H]naloxone binding was used to determine precisely the spatial organization of the patch compartment in the rat striatum and its reproducibility in different animals. Three-dimensional reconstruction of the patch network was made using maps of autoradiographic data obtained from successive coronal, sagittal or horizontal sections. The extreme rostral pole of the striatum (A 11) was characterized by a large patch territory exhibiting complex and tortuous fields with several extensions. In the intermediate part of the structure (A 9.0–10.0), about 20 serial parallel continuous patch channels running in a mediolateral axis, obliquely oriented and displaying in some cases connecting branches, could be observed. However, no channels could be distinguished in the rostrocaudal direction. More caudally, patches were rare and of small size. In addition, the laterocaudal region of the striatum was almost exclusively represented by a large matrix field. Finally, a fine discontinuous band of [ 3H]naloxone binding was seen in all sections, bordering and limiting the dorsolateral part of the striatum. The topographical and spatial distribution of the patch compartment was similar in all animals investigated. However, due to the tortuous shape and the labyrinthine organization of the patches, the precise degree of reproducibility from one animal to another could not be established. Nevertheless, the prominent patch compartment observed in the rostral pole of the striatum, the patch channels, oriented in the mediolateral axis as well as the large laterocaudal matrix field were observed in all cases. These results were compared with previous data obtained in the cat in which patch (striosome) channels oriented along a rostrocaudal axis are also observed.

Chronic morphine increases calbindin D28k in rat striatum: possible NMDA receptor involvement.

The neuronal intracellular calcium-binding protein calbindin D28k is a neurochemical marker in the striatum, normally expressed in the matrix compartment and absent from the mu-opiate receptor-enriched striosomal (patch) compartment. Because chronic treatment with morphine has been reported to increase calcium levels in synaptosomes of rat striatum, we studied effects of morphine on calbindin immunoreactivity (ir) in rat brain, using immunocytochemistry. Treatment with morphine for 5 days increased calbindin-ir in the striatal matrix, and induced intense calbindin-ir in the patch compartment. Increased calbindin-ir in patches persisted through day 14 post-morphine. Co-administration of the NMDA receptor antagonist MK-801 blocked the morphine-induced increase in calbindin-ir in patches. We suggest that chronic morphine treatment may increase calcium and calbindin levels via increased glutamatergic transmission in striatum. These findings are consistent with reports that MK-801 inhibits tolerance to morphine, and provide a possible mechanism and anatomical substrate for this inhibition.

Pattern Formation in the Developing Mammalian Forebrain: Selective Adhesion of Early but Not Late Postmitotic Cortical and Striatal Neurons within Forebrain Reaggregate Cultures

The mammalian cerebral cortex and striatum exhibit spatially restricted distributions of neurons. These neuronal collectives correlate with the age at which the neurons become postmitotic. We investigated in vitro the role of cell adhesion in the organization of like-birthdated neurons within the rat telencephalon. Specifically, the ability of neurons of similar birthdate to reassociate with one another within either striatal or cortical reaggregates was observed. Early postmitotic neurons (destined for the striatal patch compartment and cortical deep layers) or later born neurons (destined for the striatal matrix compartment and cortical superficial layers) were labeled in vivo with [3H]-thymidine or bromodeoxyuridine (Brdu) on Embryonic Day (E) 13 or 18, respectively. Two or seven days later, the striatum and cortex were separately removed, dissociated, and reaggregated in suspension cultures for 5 days. Within both striatal and cortical reaggregates, E13 [3H]thymidine-labeled cells were observed to clump together toward the center of the reaggregates. Conversely, reaggregates containing cells labeled on E18 with a single [3H]thymidine injection or with two separate Brdu injections contained labeled cells which were dispersed with respect to each other, but showed an overall distribution toward the periphery of the reaggregates. These results suggest that early, but not late, postmitotic striatal and cortical neurons selectively associate with one another within their respective structures. Dissociated embryonic striatal and cortical neurons were then coreaggregated to examine if a single adhesive mechanism is shared by both tissues. Early born neurons within these reaggregates clustered with each other regardless of tissue type. Later born cortical and striatal neurons were found dispersed in relation to each other and the early born neurons. However, the selective adhesion of early born striatal and cortical neurons is not an attribute of all early postmitotic forebrain neurons, because dissociated early born septal-basal forebrain neurons were found dispersed within reaggregates of septal-basal forebrain tissue or when cocultured with striatal tissue. These results suggest that the organization of cortical and striatal neuronal groups may depend on a common adhesive mechanism that crosses tissue-type boundaries.

Regionally specific effects of atypical antipsychotic drugs on striatal Fos expression: The nucleus accumbens shell as a locus of antipsychotic action

The mechanisms by which atypical antipsychotic drugs such as clozapine exert therapeutic effects but do not induce extrapyramidal side effects are not clear. We have examined the effects of acute administration of three antipsychotic drugs on Fos protein expression in the striatal complex. The ypical neuroleptic haloperidol was compared with the atypical agent clozapine and the putative atypical antipsychotic drug remoxipride. Haloperidol increased the number of neurons expressing Fos-like immunoreactivity in both the dorsolateral and the medial striatum and increased Fos expression in the nucleus accumbens core and shell. Clozapine increased Fos in the nucleus accumbens shell, but not in the core or two neostriatal sectors. Remoxipride significantly increased the number of Fos-like immunoreactive neurons in the medial but not the dorsolateral striatum, and increased the number of cells expressing Fos protein in the nucleus accumbens shell but not core. The remoxipride-induced increase in the number of medial striatal neurons expressing Fos was entirely attributable to a selective increase in the striatal patch compartment, whereas haloperidol increased Fos protein in neurons of both striatal compartments. These data indicate that typical and atypical antipsychotic drugs exert regionally distinct effects on striatal Fos expression, and suggest that the dorsolateral striatum may be a locus involved in the genesis of extrapyramidal side effects. All three antipsychotic drugs increased Fos expression in the shell of the nucleus accumbens. The shell of the nucleus accumbens may be a site of antipsychotic action.

Neonatal 6-OHDA lesions differentially affect striatal D1 and D2 receptors.

Lesions to the dopamine (DA) system in early postnatal development have different behavioral consequences compared to lesions made in adulthood. Intrastriatal injections of the neurotoxin 6-hydroxydopamine (6-OHDA) on the day of birth (PO) or postnatal day 1 (P1) produce a selective supersensitivity to D1 receptor agonists and a subsensitivity to D1 antagonists (Neal and Joyce, 1991a). In this paper, we describe the long-term effects of early DA loss on DA receptor regulation. Pups received bilateral intrastriatal injections of the neurotoxin 6-OHDA (4 micrograms per striatum) on PO or P1. Adult rats were killed at 90 days of age and the brains were processed for quantitative autoradiography (QAR) or tyrosine hydroxylase (TH) immunocytochemistry. Cohorts were tested for the behavioral responses to the selective D1 receptor agonist SKF38393 (10 mg/kg). Neonatally lesioned rats exhibited increases in abnormal perioral movements in response to D1 receptor stimulation. There was a heterogenous and patchy loss (40-50%) of [3H]mazindol binding to high-affinity DA uptake sites (a marker of DA terminal density) and a similar loss of TH-like immunoreactivity within the striata of the neonatally lesioned rats. There was also a reduction in the number of mu-opioid receptor patches (labelled with [3H]naloxone), a marker for the striatal patch compartment, and a similar patchy loss of D1 binding sites (labeled with [3H]SCH23390). The binding of [3H]spiroperidol to D2 sites was not altered. This is in contrast to the changes observed following adult 6-OHDA lesions, wherein there is a significant increase in the number of D2 binding sites (Joyce, 1991a,b). The results are discussed with respect to the behavioral consequences of neonatal lesions and the differences between neonatal and adult lesions.

Ontogeny of gene expression of adenosine A2 receptor in the striatum: early localization in the patch compartment.

The ontogeny of adenosine A2 receptor mRNA and adenosine A2 binding sites distributions was studied by in situ hybridization histochemistry and receptor autoradiography in pre- and post-natal rat striatum, postnatal dog striatum, and a human fetus striatum and compared to that of dopamine D1 and mu opiate receptors. The early postnatal striatum demonstrated heterogeneous distributions of adenosine A2 receptor mRNA and adenosine A2 binding sites with patches of dense labeling corresponding to dopamine D1 and mu opiate receptors enriched zones. This patchy pattern evolved to the homogeneous distribution observed in the adult. The higher intensity of adenosine A2 receptor mRNA enriched patches correspond at the microscopical level to a higher density of labeled neurons in the patches areas and also to a higher level of expression per labeled patches neuron than in the matrix ones. This demonstrates for the first time that differences in patch/matrix receptor density is at least partly linked to different levels of receptor gene expression.

Embryonic lesions of the substantia nigra prevent the patchy expression of opiate receptors, but not the segregation of patch and matrix compartment neurons, in the developing rat striatum

Unilateral lesions of the substantia nigra on embryonic day 19 prevent the development of the normal patchy distribution of opiate receptors in the ipsilateral rat striatum. Independent, early and permanent labelling of patch compartment neurons in the same brains on embryonic day 14 with [ 3H]thymidine revealed that the substantia nigra lesions did not prevent the aggregation of early born neurons into patches, but rather blocked the normal expression of one phenotype (dense opiate receptor binding) of these patches. Thus, early nigrostriatal connections may not be critical for the fundamental patch/matrix compartmentation of the striatum, but may be important in the maturation of phenotypic markers of these compartments.

Subsets of neurotensin-immunoreactive neurons revealed following antagonism of the dopamine-mediated suppression of neurotensin immunoreactivity in the rat striatum

The distribution of neurotensin-immunoreactive structures in the rat striatum was evaluated after blockade of dopamine neurotransmission by drugs that act presynaptically (6-hydroxydopamine, reserpine) and postsynaptically, preferentially at the D 2 (eticlopride, haloperidol) and D 1 [( R)-(+)-8-chloro-2,3,4,5-tetrahydro-3-methyl-5-phenyl-1H-3-benzazepin-7-ol, SCH-23390] receptor sites. Calbindin-D (mol. wt 28,000) immunoreactivity was used to delineate patch (striosome) and matrix in the caudate-putamen and core and shell in the nucleus accumbens. Antagonism at the D 2 dopamine receptor and 6-hydroxydopamine lesions caused dense axonal immunoreactivity and moderate numbers of neurotensin-immunoreactive neurons to be distributed preferentially in the matrix of the caudate-putamen. D 1 receptor antagonism was significantly less effective at eliciting neurotensin-immuno-reactive neurons in the caudate-putamen. Reserpine or co-administration of the D 1 and D 2 receptor antagonists produced many neurotensin-immunoreactive neurons in both striatal compartments throughout the caudate-putamen and dense axonal neurotensin immunoreactivity in the medial patch compartment. To varying degrees, with SCH-23390 being least effective and reserpine most effective, all of the drug treatments elicited neurotensin immunoreactivity in neurons in the olfactory tubercle, rostral nucleus accumbens, accumbal shell and ventrolateral caudate-putamen, i.e. most of the ventral striatum.